SB225002-DataSheet-生命科學試劑-MedChemExpress

Hotline:400-820-3792Inhibitors?Agonists?ScreeningLibrarieswww.MedChemESB225002Cat.No.:HY-16711CASNo.:182498-32-4分?式:C??H??BrN?O?分?量:352.14作?靶點:CXCR作?通路:GPCR/GProtein;Immunology/Inflammation儲存?式:4°C,storedundernitrogen*Insolvent:-80°C,6months;-20°C,1month(storedundernitrogen)溶解性數(shù)據(jù)體外實驗DMSO:≥100mg/mL(283.98mM)掃描?維碼，*"≥"meanssoluble,butsaturationunknown.運?溶解?案計算器獲得適合您實驗體系的溶解?案MassSolvent1mg5mg10mgConcentration制備儲備液1mM2.8398mL14.1989mL28.3978mL5mM0.5680mL2.8398mL5.6796mL10mM0.2840mL1.4199mL2.8398mL請根據(jù)產(chǎn)品在不同溶劑中的溶解度，選擇合適的溶劑配制儲備液，并請注意儲備液的保存?式和期限。體內實驗請根據(jù)您的實驗動物和給藥?式選擇適當?shù)娜芙?案。以下溶解?案都請先按照InVitro?式配制澄的儲備液，再依次添加助溶劑：為保證實驗結果的可靠性，澄的儲備液可以根據(jù)儲存條件，適當保存；體內實驗的?作液，建議您現(xiàn)?現(xiàn)配，當天使?；以下溶劑前顯?的百分?指該溶劑在您配制終溶液中的體積占?；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的?式助溶1.請依序添加每種溶劑：10%DMSO40%PEG3005%Tween-8045%salineSolubility:≥2.5mg/mL(7.10mM);Clearsolution此?案可獲得≥2.5mg/mL(7.10mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到400μLPEG300中，混合均勻；向上述2.體系中加?50μLTween-80，混合均勻；然后繼續(xù)加?450μL?理鹽?定容?1mL。請依序添加每種溶劑：10%DMSO90%(20%SBE-β-CDinsaline)1/4www.MedChemEwww.MedChemESolubility:≥2.5mg/mL(7.10mM);Clearsolution此?案可獲得≥2.5mg/mL(7.10mM，飽和度未知)的澄溶液。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到900μL20%的SBE-β-CD?理鹽??溶3.液中，混合均勻。請依序添加每種溶劑：10%DMSO90%cornoilSolubility:≥2.5mg/mL(7.10mM);Clearsolution此?案可獲得≥2.5mg/mL(7.10mM，飽和度未知)的澄溶液，此?案不適?于實驗周期在半個?以上的實驗。以1mL?作液為例，取100μL25.0mg/mL的澄DMSO儲備液加到900μL??油中，混合均勻。BIOLOGICALACTIVITY?物活性SB225002?種有效的選擇性CXCR2?肽拮抗劑，抑制125I-IL-8和CXCR2結合的IC50為22nM。IC50&Target125I-IL-8-CXCR222nM(IC50,inCHOcellmembrane)體外研究SB225002(SB225002)isanantagonistof125I-IL-8bindingtoCXCR2withanIC50=22nM.SB225002shows>150-foldselectivityoverCXCR1andfourother7-TMRstested.SB225002isapotentantagonistofrabbitCXCR2,inhibitingrabbitPMNchemotaxisinresponsetooptimalconcentrationsofhumanIL-8orGROα(IC50valuesof30and70nM,respectively.Inthesecells(PMN,HL60,CXCR1-RBL-2H3),SB225002producesaconcentration-dependentinhibitionofbothIL-8-andGROα-mediatedcalciummobilizationwithIC50valuesof8and10nM,respectively.In3ASubEcellsstablytransfectedwithCXCR2,SB225002dose-dependentlyinhibitscalciummobilizationinducedbybothGROαandIL-8,withIC50valuesof20and40nM,respectively[1].WHCO1cellstreatedwithSB225002exhibitsa40%reductionincellproliferation.BlockingCXCR2signalinginWHCO1cellswith400nMSB225002(SB225002)significantlydecreasescellproliferationby~40%to50%[2].體內研究SB225002(SB225002)selectivelyblocksIL-8-inducedneutrophilmarginationinrabbits[1].CXCR2isblockedusingtheselectiveantagonistSB225002(2mg/kg)orneutralizingCXCR2antiserum.TheCXCR2antagonistSB225002decreasesneutrophilcountsinischemichemispheresofApoE?/?miceonWesterndietandwildtypemiceonnormaldiet[3].SB225002significantlyattenuatesmicroglialactivationandBBBdamage,increasesmyelination,andreducesastrogliosisinthewhitematterafterLPS-sensitizedHI[4].PROTOCOLKinaseAssay[1]CHO-CXCR1andCHO-CXCR2membranesareprepared.Assaysareperformedin96-wellmicrotiterplateswherethereactionmixturecontained1.0μg/mLmembraneproteinin20mMBis-Tris-propane,pH8.0,with1.2mMMgSO4,0.1mMEDTA,25mMNaCl,and0.03%CHAPSandSB225002(10mMstockinMe2SO)addedattheindicatedconcentrations,thefinalMe2SOconcentrationis<1%understandardbindingconditions.Bindingisinitiatedbyadditionof0.25nM125I-IL-8(2,200Ci/mmol).After1-hincubationatroomtemperaturetheplateisharvestedusingaTomtec96-wellharvesterontoaglassfiberfiltermatblockedwith1%polyethyleneimine,0.5%BSAandwashedthreetimeswith25mMNaCl,10mMTris?HCl,1mMMgSO4,2/4www.MedChemEwww.MedChemE0.5mMEDTA,0.03%CHAPS,pH7.4.Thefilterisdried,sealedinasamplebagcontaining10mLofWallac205Betaplateliquidscintillationfluid,andcountedwithaWallac1205Betaplateliquidscintillationcounter[1].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.CellAssay[2]ThreeesophagealsquamouscellcarcinomacelllinesWHCO1,WHCO5,andWHCO6originallyestablishedfromsurgicalbiopsiesofprimaryesophagealsquamouscellcarcinomasareculturedinDMEMcontaining10%FCSat37°Cinahumidifiedatmosphereof5%CO2.MTTassaysarecarriedoutusingtheCellProliferationkit.Briefly,1.5×103cellsareplatedin96-wellplatesinafinalvolumeof180μLDMEMperwell.SB225002(400nM)isaddedtocellsand0.001%DMSO(solvent)isaddedasacontrol.Aftertheindicatedincubationperiod,18μLoftheMTTlabelingreagent(finalconcentration0.5mg/mL)isaddedtoeachwellandincubatedfor4hoursinahumidifiedatmosphere.Onehundredeightymicrolitersofthesolubilizationsolutionareaddedtoeachwellandtheplatesareleftovernightat37°C.Thespectrophotometricabsorbanceofsamplesismeasuredat595nmusingamicrotiterplatereader[2].MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.AnimalMice[3]Administration[3][4]Male7-8weeksoldwildtype(C57BL/6J,Harlan)andApoE?/?mice,whicharegeneratedonthesameC57BL/6background,areeitherfedwithanormalchoworacholesterolrichchowfor6weeksandsubmittedto20minofleft-sidedmiddlecerebralarteryocclusion(MCAO)orshamsurgery.Animalsarerandomlyattributedtotreatmentparadigms,andexperimentersareblindedatallstagesofinterventionsanddataanalysis.TheselectiveCXCR2antagonistSB225002(2mg/kg)orvehicle(1%DMSOinPBS)isinjectedintraperitoneally(i.p.)at0,24and48hourspost-ischemia.Inotherexperiments,CXCR2isspecificallyblockedbyi.p.injectionofaneutralizingrabbitanti-CXCR2serum(300μL)at0hours,24hoursand48hourspost-ischemia.Inthelatterstudies,normalrabbitserum(NRS)servedascontrol.Insomeexperiments,neutrophilsaredepletedbyi.p.injectionof200μganti-mouseLy6G24hoursbeforeand24hoursafterischemia.Intheseexperiments,200μgofanisotypecontrolantibodyisdeliveredascontrol.Rats[4]Inthisstudy,10-12Sprague-Dawleyratpupsperdamareused.Thepupsreceiveintraperitonealinjections

ofSB225002(1or3mg/kg,dilutedinNScontaining0.33%Tween80)orvehicle(NSsolutioncontaining0.33%Tween80)30minbeforelipopolysaccharide(LPS)administrationandimmediatelyafterhypoxic

ischemia(HI).Thepupsarerandomlyassignedtofourgroups:control(pupsunexposedtoLPSorHI,N=14),

vehicle(NSinjections30minbeforeLPSadministrationandimmediatelyafterHI,N=18),andSB-1(1mg/kg,

N=14)andSB-3(3mg/kg,N=18)(SB225002injections30minbeforeLPSadministrationandimmediately

afterHI).MCEhasnotindependentlyconfirmedtheaccuracyofthesemethods.Theyareforreferenceonly.戶使?本產(chǎn)品發(fā)表的科研?獻?NatCommun.2020Feb28;11(1):1126.?CancerRes.2018Oct1;78(19):5586-5599.?BioactMater.6(2021)2039-2057.?JAutoimmun.2018May;89:30-40.?JCellPhysiol.2021Apr;236(4):3114-3128.3/4www.MedChemEwww.MedChemESeemorecustomervalidationsonwww.MedChemEREFERENCES[1].WhiteJR,etal.Identificationofapotent,selectivenon-peptideCXCR2antagonistthatinhibitsinterleukin-8-inducedneutrophilmigration.JBiolChem.1998Apr24;273(17):10095-8.[2].WangB,etal.Agrowth-relatedoncogene/CXCchemokinereceptor2autocrineloopcontributestocellularproliferationinesophagealcancer.CancerRes.20